Western Blot was performed using Anti-ACE Recombinant Rabbit Monoclonal Antibody (JM59-32) (Product # MA5-35099) and a 200 kDa band corresponding to ACE was observed except in Rat Liver and Rat Heart (DOI: 10.1161/HYPERTENSIONAHA.111.182428). Expression of ACE was increased upon serum starvation of HUVEC and treatment with PMA, also decreased upon treatment with LPS (DOI: 10.1161/HYPERTENSIONAHA.111.182428, 10.1007/s11626-013-9707-0). Tissue extracts (30 µg lysate) of Mouse Aorta (Lane 1), Mouse Lung (Lane 2), Mouse Liver (Lane 3), Mouse Heart (Lane 4), Rat Aorta (Lane 5), Rat Lung (Lane 6), Rat Liver (Lane 7), Rat Heart (Lane 8) (Fig a), membrane enriched extracts of HUVEC (Lane 1), Serum starved HUVEC treated with PMA (24 hours, and 100 ng/mL for 24 hours) (Lane 2), HUVEC (Lane 3), HUVEC treated with LPS (0.5 µg/mL for 24 hours) (Lane 4) (Fig b) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The Blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752).